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      Effects of leucine-enriched essential amino acid and whey protein bolus dosing upon skeletal muscle protein synthesis at rest and after exercise in older women

      research-article
      a , 1 , a , 1 , a , a , a , a , a , a , b , a , a , a , a , a ,
      Clinical Nutrition (Edinburgh, Scotland)
      Elsevier
      Muscle protein synthesis, Leucine, Ageing, Low dose amino acid supplementation, Exercise, Human metabolism, MPS, muscle protein synthesis, EAA, essential amino acids, WP, whey protein, LEAA, leucine enriched essential amino acids, LBF, leg blood flow, MBF, microvascular blood flow, BMI, body mass index, SMI, skeletal muscle index, 1-RM, 1 repetition maximum, PVDF, polyvinylidene difluoride, TBST, tris buffered saline/Tween 20, CEUS, contrast enhanced ultrasound

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          Summary

          Background & aims

          Impaired anabolic responses to nutrition and exercise contribute to loss of skeletal muscle mass with ageing (sarcopenia). Here, we tested responses of muscle protein synthesis (MPS), in the under represented group of older women, to leucine-enriched essential amino acids (EAA) in comparison to a large bolus of whey protein (WP).

          Methods

          Twenty-four older women (65 ± 1 y) received ( N = 8/group) 1.5 g leucine-enriched EAA supplements (LEAA_1.5), 6 g LEAA (LEAA_6) in comparison to 40 g WP. A primed constant I.V infusion of 13C 6-phenylalanine was used to determine MPS at baseline and in response to feeding (FED) and feeding-plus-exercise (FED-EX; 6 × 8 unilateral leg extensions; 75%1-RM). We quantified plasma insulin/AA concentrations, leg femoral blood flow (LBF)/muscle microvascular blood flow (MBF), and anabolic signalling via immunoblotting.

          Results

          Plasma insulineamia and EAAemia were greater and more prolonged with WP than LEAA, although LEAA_6 peaked at similar levels to WP. Neither LEAA or WP modified LBF or MBF. FED increased MPS similarly in the LEAA_1.5, LEAA_6 and WP ( P < 0.05) groups over 0–2 h, with MPS significantly higher than basal in the LEAA_6 and WP groups only over 0–4 h. However, FED-EX increased MPS similarly across all the groups from 0 to 4 h ( P < 0.05). Only p-p70S6K1 increased with WP at 2 h in FED ( P < 0.05), and at 2/4 h in FED-EX ( P < 0.05).

          Conclusions

          In conclusion, LEAA_1.5, despite only providing 0.6 g of leucine, robustly (perhaps maximally) stimulated MPS, with negligible trophic advantage of greater doses of LEAA or even to 40 g WP. Highlighting that composition of EAA, in particular the presence of leucine rather than amount is most crucial for anabolism.

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          Most cited references29

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          Coordinated collagen and muscle protein synthesis in human patella tendon and quadriceps muscle after exercise.

          We hypothesized that an acute bout of strenuous, non-damaging exercise would increase rates of protein synthesis of collagen in tendon and skeletal muscle but these would be less than those of muscle myofibrillar and sarcoplasmic proteins. Two groups (n = 8 and 6) of healthy young men were studied over 72 h after 1 h of one-legged kicking exercise at 67% of maximum workload (W(max)). To label tissue proteins in muscle and tendon primed, constant infusions of [1-(13)C]leucine or [1-(13)C]valine and flooding doses of [(15)N] or [(13)C]proline were given intravenously, with estimation of labelling in target proteins by gas chromatography-mass spectrometry. Patellar tendon and quadriceps biopsies were taken in exercised and rested legs at 6, 24, 42 or 48 and 72 h after exercise. The fractional synthetic rates of all proteins were elevated at 6 h and rose rapidly to peak at 24 h post exercise (tendon collagen (0.077% h(-1)), muscle collagen (0.054% h(-1)), myofibrillar protein (0.121% h(-1)), and sarcoplasmic protein (0.134% h(-1))). The rates decreased toward basal values by 72 h although rates of tendon collagen and myofibrillar protein synthesis remained elevated. There was no tissue damage of muscle visible on histological evaluation. Neither tissue microdialysate nor serum concentrations of IGF-I and IGF binding proteins (IGFBP-3 and IGFBP-4) or procollagen type I N-terminal propeptide changed from resting values. Thus, there is a rapid increase in collagen synthesis after strenuous exercise in human tendon and muscle. The similar time course of changes of protein synthetic rates in different cell types supports the idea of coordinated musculotendinous adaptation.
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            Coomassie staining as loading control in Western blot analysis.

            In Western blotting, immunodetection of housekeeping proteins is routinely performed to detect differences in electrophoresis loading. The present work describes a much faster and simpler protein staining method, which is compatible with ordinary blocking conditions. In addition, the method can be used after immunodetection with superior linearity compared to ordinary staining methods. After immunoblotting and staining, protein bands can be further identified using peptide mass fingerprinting.
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              Leucine supplementation of a low-protein mixed macronutrient beverage enhances myofibrillar protein synthesis in young men: a double-blind, randomized trial.

              Leucine is a key amino acid involved in the regulation of skeletal muscle protein synthesis. We assessed the effect of the supplementation of a lower-protein mixed macronutrient beverage with varying doses of leucine or a mixture of branched chain amino acids (BCAAs) on myofibrillar protein synthesis (MPS) at rest and after exercise. In a parallel group design, 40 men (21 ± 1 y) completed unilateral knee-extensor resistance exercise before the ingestion of 25 g whey protein (W25) (3.0 g leucine), 6.25 g whey protein (W6) (0.75g leucine), 6.25 g whey protein supplemented with leucine to 3.0 g total leucine (W6+Low-Leu), 6.25 g whey protein supplemented with leucine to 5.0 g total leucine (W6+High-Leu), or 6.25 g whey protein supplemented with leucine, isoleucine, and valine to 5.0 g total leucine. A primed continuous infusion of l-[ring-(13)C6] phenylalanine with serial muscle biopsies was used to measure MPS under baseline fasted and postprandial conditions in both a rested (response to feeding) and exercised (response to combined feeding and resistance exercise) leg. The area under the blood leucine curve was greatest for the W6+High-Leu group compared with the W6 and W6+Low-Leu groups (P < 0.001). In the postprandial period, rates of MPS were increased above baseline over 0-1.5 h in all treatments. Over 1.5-4.5 h, MPS remained increased above baseline after all treatments but was greatest after W25 (∼267%) and W6+High-Leu (∼220%) treatments (P = 0.002). A low-protein (6.25 g) mixed macronutrient beverage can be as effective as a high-protein dose (25 g) at stimulating increased MPS rates when supplemented with a high (5.0 g total leucine) amount of leucine. These results have important implications for formulations of protein beverages designed to enhance muscle anabolism. This trial was registered at clinicaltrials.gov as NCT 1530646.
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                Author and article information

                Contributors
                Journal
                Clin Nutr
                Clin Nutr
                Clinical Nutrition (Edinburgh, Scotland)
                Elsevier
                0261-5614
                1532-1983
                1 December 2018
                December 2018
                : 37
                : 6Part A
                : 2011-2021
                Affiliations
                [a ]MRC/ARUK Centre of Excellence for Musculoskeletal Ageing Research, National Institute for Health Research Nottingham Biomedical Research Centre, University of Nottingham, Derby DE22 3DT, UK
                [b ]Ajinomoto Co., Inc., Tokyo 104-8315, Japan
                Author notes
                []Corresponding author. MRC–ARUK Centre for Musculoskeletal Ageing Research, National Institute for Health Research Nottingham Biomedical Research Centre, School of Medicine, University of Nottingham, Derby DE22 3DT, UK. Fax: +44 1332 724727. philip.atherton@ 123456nottingham.ac.uk
                [1]

                Authors contributed equally.

                Article
                S0261-5614(17)31340-7
                10.1016/j.clnu.2017.09.008
                6295981
                29031484
                9804967c-47d2-453f-a85a-c263ea1802b8
                © 2017 The Authors

                This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

                History
                : 9 April 2017
                : 19 September 2017
                Categories
                Article

                muscle protein synthesis,leucine,ageing,low dose amino acid supplementation,exercise,human metabolism,mps, muscle protein synthesis,eaa, essential amino acids,wp, whey protein,leaa, leucine enriched essential amino acids,lbf, leg blood flow,mbf, microvascular blood flow,bmi, body mass index,smi, skeletal muscle index,1-rm, 1 repetition maximum,pvdf, polyvinylidene difluoride,tbst, tris buffered saline/tween 20,ceus, contrast enhanced ultrasound

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